Changes of multiple autophagy pathways in dopaminergic cells under oxidative stress
10.3760/cma.j.issn.1671-8925.2014.03.009
- VernacularTitle:氧化应激条件下多巴胺能细胞自噬的变化
- Author:
Lan WANG
1
;
Wei SHEN
;
Yanwei SU
Author Information
1. 华中科技大学同济医学院附属普爱医院神经内科
- Keywords:
Oxidative stress;
Autophagy;
PC12 cells;
Rotenone
- From:
Chinese Journal of Neuromedicine
2014;13(3):261-264
- CountryChina
- Language:Chinese
-
Abstract:
Objective To study the activity of macroautophagy and chaperone-medicated antophagy (CMA) in dopaminergic cells under oxidative stress and their possible roles in neurodegeneration.Methods PC 12 cells were treated with various concentrations of rotenone (0,20,100 and 500 nmol/L).The morphological characteristics of autophagosomes were observed by transmission electron microscopy (TEM),and macroautophagic activity was determined by detecting light chain (LC3) expression of microtubule-associated protein by Western blotting and monodansylcadaverine (MDC) immunofluorescence.Molecular chaperone mediated autophagy was estimated by the expressions of lysosomal-associated membrane protein 2α (Lamp-2α),heat shock cognate 70 (Hsc70) and heat shock protein 90 (Hsp90),which were detected by Western blotting.Results In control group (0 nmol/L rotenone),TEM showed that the cells presented a low autophagic activity with less autophagic vacuoles; in rotenone treatment groups,the autophagosomes with different forms were significantly increased,and the phenomenon ofautophagic mitochondria was observed.A large of vacuole-like bodies were found in the cytoplasm of the cells treated with high concentration of rotenone.The MDC immunofluorescence showed that after the treatment of rotenone,the red fluorescence showed punctate distribution,which was also significantly enhanced with the increasing of the concentrations of rotenone.Western blotting showed dose-dependent expressions of LC3-Ⅰ and LC3-Ⅱ.As compared with control cells,the expressions ofLamp-2α,Hsc70 and Hsp90 in rotenone treatment groups were also increased significantly (P＜0.05),but didn't show statistical difference between each two rotenone treatment groups (P＞0.05).Conclusions The macroautophagy and autophagy mediated by molecular chaperone are both activated in PC12 cells under oxidative stress conditions.And the increased oxidative stress results in an enhanced activity of macroautophagy,while the activity of CMA is saturated.
